Amyotrophic lateral sclerosis, commonly referred to as Lou Gehrig's disease, is characterized by selective, premature degeneration and death of motor neurons in the motor cortex, brain stem and spinal cord. The loss of motor neurons causes progressive muscle paralysis ultimately leading to death from respiratory failure. Approximately 90% of all amyotrophic lateral sclerosis cases are sporadic amyotrophic lateral sclerosis, without a family history of the disease, and the other approximately 10% of cases are cases of familial amyotrophic lateral sclerosis. Despite significant efforts to identify risk factors and potential susceptibility genes, the etiology of sporadic amyotrophic lateral sclerosis remains largely unknown.
Various rodent models carrying dominant mutations of the human superoxide dismutase (SOD1) that is causative in about 20% of familial amyotrophic lateral sclerosis cases, have been instrumental to model motor neuron toxicity in amyotrophic lateral sclerosis. These models have demonstrated that not only motor neurons, but also non-neuronal cell types including microglia and astrocytes play a significant role in disease onset and progression. Recent studies have identified astrocytes as mediators of motor neuron death in amyotrophic lateral sclerosis by a yet undetermined cytotoxic mechanism. Insight into the mechanisms underlying the acquisition of this toxic function by amyotrophic lateral sclerosis astrocytes is pertinent for the development of successful therapies for amyotrophic lateral sclerosis.
Accordingly, the present inventors have discovered the mechanism underlying the acquisition of this toxic function by amyotrophic lateral sclerosis astrocytes, and have used this knowledge to develop therapies for amyotrophic lateral sclerosis. The pharmaceutical compositions, methods and uses, and kits described herein can be used to treat sporadic or familial amyotrophic lateral sclerosis.
Several embodiments of the invention are described by the following enumerated clauses:
1. A method for treating a patient with amyotrophic lateral sclerosis by decreasing the expression of a cytoplasmic granule toxin in astrocytes of the patient, the method comprising the step of                administering to the patient a composition comprising an effective amount of a compound that decreases the expression the cytoplasmic granule toxin in the astrocytes of the patient.        
2. The method of clause 1 wherein the cytoplasmic granule toxin is a perforin.
3. The method of clause 2 wherein the perforin is perforin 1.
4. The method of clause 1 wherein the cytoplasmic granule toxin is a granzyme.
5. The method of clause 4 wherein the granzyme is granzyme B.
6. The method of any one of clauses 1 to 5 wherein the decreased expression of the cytoplasmic granule toxin results in an effect on motor neurons in the patient selected from the group consisting of an increase in the number of motor neurons, a decrease in soma atrophy, and an increase in neurite length after administration of the compound.
7. The method of any one of clauses 1 to 6 wherein the compound is selected from the group consisting of a drug, a peptide, and a nucleic acid.
8. The method of clause 7 wherein the compound is a nucleic acid.
9. The method of clause 8 wherein the nucleic acid functions by RNA interference or is an antisense RNA molecule.
10. The method of clause 8 wherein the nucleic acid is selected from the group consisting of an siRNA, an miRNA, and an shRNA.
11. The method of clause 10 wherein the nucleic acid is an shRNA.
12. The method of clause 8 wherein the nucleic acid is delivered to the patient in a bacterial vector or in a viral vector.
13. The method of clause 8 wherein the nucleic acid has the sequence of SEQ ID NO: 1.
14. The method of clause 8 wherein the nucleic acid has the sequence of SEQ ID NO: 2.
15. The method of any one of clauses 1 to 14 wherein the amyotrophic lateral sclerosis is sporadic amyotrophic lateral sclerosis.
16. The method of any one of clauses 1 to 14 wherein the amyotrophic lateral sclerosis is familial amyotrophic lateral sclerosis.
17. The method of any one of clauses 1 to 16 wherein the amount of the compound is in the range of about 1 ng/kg of patient body weight to about 1 mg/kg of patient body weight.
18. The method of any one of clauses 1 to 17 wherein the amount of the compound is in the range of about 1 ng/kg of patient body weight to about 500 ng/kg of patient body weight.
19. The method of any one of clauses 1 to 18 wherein the amount of the compound is in the range of about 1 ng/kg of patient body weight to about 100 ng/kg of patient body weight.
20. The method of any one of clauses 1 to 19 wherein the composition further comprises a carrier, an excipient, or a diluent, or a combination thereof.
21. The method of clause 20 wherein the composition comprises a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is a liquid carrier.
22. The method of clause 21 wherein the liquid carrier is selected from the group consisting of saline, glucose, alcohols, glycols, esters, amides, and a combination thereof.
23. The method of any one of clauses 1 to 22 wherein the composition is administered in a single-dose or a multiple-dose regimen.
24. A method for treating amyotrophic lateral sclerosis by increasing MHC class I expression in motor neurons of a patient, the method comprising the step of                administering to the patient a composition comprising an effective amount of a compound that increases the expression of MHC class I in the motor neurons of the patient.        
25. The method of clause 24 wherein the increased expression of the MHC class I results in an effect on motor neurons in the patient selected from the group consisting of an increase in the number of motor neurons, a decrease in soma atrophy, and an increase in neurite length after administration of the compound.
26. The method of any one of clauses 24 to 25 wherein the compound is selected from the group consisting of a drug, a peptide, and a nucleic acid.
27. The method of clause 26 wherein the compound is a nucleic acid.
28. The method of clause 27 wherein the nucleic acid is delivered to the patient in a bacterial vector or in a viral vector.
29. The method of clause 28 wherein the vector is a viral vector.
30. The method of clause 29 wherein the vector is selected from the group consisting of a lentiviral vector and an adenovirus vector.
31. The method of clause 30 wherein the nucleic acid has the sequence of SEQ ID NO: 3.
32. The method of clause 31 wherein the nucleic acid encodes the histocompatibility complex H2K.
33. The method of any one of clauses 24 to 32 wherein the amyotrophic lateral sclerosis is sporadic amyotrophic lateral sclerosis.
34. The method of any one of clauses 24 to 32 wherein the amyotrophic lateral sclerosis is familial amyotrophic lateral sclerosis.
35. The method of any one of clauses 24 to 34 wherein the amount of the compound is in the range of about 1 ng/kg of patient body weight to about 1 mg/kg of patient body weight.
36. The method of any one of clauses 24 to 35 wherein the amount of the compound is in the range of about 1 ng/kg of patient body weight to about 500 ng/kg of patient body weight.
37. The method of any one of clauses 24 to 36 wherein the amount of the compound is in the range of about 1 ng/kg of patient body weight to about 100 ng/kg of patient body weight.
38. The method of any one of clauses 24 to 37 wherein the composition further comprises a carrier, an excipient, or a diluent, or a combination thereof.
39. The method of clause 38 wherein the composition comprises a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is a liquid carrier.
40. The method of clause 39 wherein the liquid carrier is selected from the group consisting of saline, glucose, alcohols, glycols, esters, amides, and a combination thereof.
41. The method of any one of clauses 24 to 40 wherein the composition is administered in a single-dose or a multiple-dose regimen.
42. A pharmaceutical composition comprising a dosage form of a compound effective to decrease the expression a cytoplasmic granule toxin in the astrocytes of a patient with amyotrophic lateral sclerosis.
43. The composition of clause 42 wherein the compound is selected from the group consisting of a drug, a peptide, and a nucleic acid.
44. The composition of clause 43 wherein the compound is a nucleic acid.
45. The composition of clause 44 wherein the nucleic acid is selected from the group consisting of siRNA, an miRNA, and an shRNA.
46. The composition of clause 44 wherein the compound is an antisense RNA molecule.
47. The composition of clause 45 wherein the nucleic acid is an shRNA.
48. The composition of clause 44 wherein the nucleic acid has the sequence of SEQ ID NO: 1.
49. The composition of clause 44 wherein the nucleic acid has the sequence of SEQ ID NO: 2.
50. The composition of any one of clauses 42 to 49, wherein the composition further comprises one or more carriers, diluents, or excipients, or a combination thereof.
51. The composition of clause 50 wherein the composition comprises a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is a liquid carrier.
52. The composition of clause 51 wherein the liquid carrier is selected from the group consisting of saline, glucose, alcohols, glycols, esters, amides, and a combination thereof.
53. The composition of any one of clauses 42 to 52 wherein the purity of the compound is at least 98% based on weight percent.
54. The composition of any one of clauses 42 to 53 wherein the composition is in an ampoule or a sealed vial.
55. The composition of any one of clauses 42 to 50 or 53 to 54 in the form of a reconstitutable lyophilizate.
56. A pharmaceutical composition comprising a dosage form of a compound effective to increase the expression of MHC class I in the motor neurons of a patient with amyotrophic lateral sclerosis.
57. The composition of clause 56 wherein the compound is selected from the group consisting of a drug, a peptide, and a nucleic acid.
58. The composition of clause 56 wherein the compound is a nucleic acid.
59. The composition of clause 58 wherein the nucleic acid has the sequence of SEQ ID NO: 3.
60. The composition of any one of clauses 56 to 59 wherein the composition further comprises one or more carriers, diluents, or excipients, or a combination thereof.
61. The composition of clause 60 wherein the composition comprises a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is a liquid carrier.
62. The composition of clause 61 wherein the liquid carrier is selected from the group consisting of saline, glucose, alcohols, glycols, esters, amides, and a combination thereof.
63. The composition of any one of clauses 56 to 62 wherein the purity of the compound is at least 98% based on weight percent.
64. The composition of any one of clauses 56 to 63 wherein the composition is in an ampoule or a sealed vial.
65. The composition of any one of clauses 56 to 60 or 63 to 64 in the form of a reconstitutable lyophilizate.
66. The method or pharmaceutical composition of any one of clauses 1 to 65 wherein the composition is in a dosage form selected from the group consisting of an inhalation dosage form, an oral dosage form, and a parenteral dosage form.
67. The method or pharmaceutical composition of clause 66 wherein the parenteral dosage form is selected from the group consisting of an intradermal dosage form, a subcutaneous dosage form, an intramuscular dosage form, an intraperitoneal dosage form, an intravenous dosage form, and an intrathecal dosage form.
68. The composition of clause 55 or 65 in the form of a lyophilizate.
69. The composition of any one of clauses 42 to 50, 53 to 60, or 63 to 65 in the form of a solid.
70. A kit comprising a sterile vial, the composition of any one of clauses 56 to 69, and instructions for use describing use of the composition for treating a patient with amyotrophic lateral sclerosis.
71. The kit of clause 70 wherein the compound or composition is in the form of a reconstitutable lyophilizate.
72. The kit of clause 70 or 71 wherein the dose of the compound is in the range of 1 to 5 μg/kg of patient body weight.
73. The kit of any one of clauses 70 to 72 wherein the purity of the compound is at least 99% based on weight percent.
74. The kit of any one of clauses 70 to 73 wherein the compound or the composition is in a parenteral dosage form.
75. The kit of clause 74 wherein the parenteral dosage form is selected from the group consisting of an intradermal dosage form, a subcutaneous dosage form, an intramuscular dosage form, an intraperitoneal dosage form, an intravenous dosage form, and an intrathecal dosage form.
76. The kit of any one of clauses 70 to 75 wherein the composition further comprises a pharmaceutically acceptable carrier.
77. The kit of clause 76 wherein the pharmaceutically acceptable carrier is a liquid carrier selected from the group consisting of saline, glucose, alcohols, glycols, esters, amides, and a combination thereof.
78. Use of the composition of any one of clauses 42 to 69 for the manufacture of a medicament for treating amyotrophic lateral sclerosis.
79. The pharmaceutical composition of any one of clauses 42 to 69 for use in treating amyotrophic lateral sclerosis.
80. The method of any one of clauses 1 to 23 wherein the cytoplasmic granule toxin is not SOD1.
81. The method of any one of clauses 1 to 23 wherein the cytoplasmic granule toxin is not an antioxidant cytoplasmic granule toxin.
82. The composition of any one of clauses 42 to 55 wherein the cytoplasmic granule toxin is not SOD1.
83. The composition of any one of clauses 42 to 55 wherein the cytoplasmic granule toxin is not an antioxidant cytoplasmic granule toxin.